LOCUS       sequence-124607-        1050 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transient expression of a minimal sgRNA targeting the GLuc reporter,
            in mammalian cells. U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1050)
  AUTHORS   Shechner DM, Hacisuleyman E, Younger ST, Rinn JL
  TITLE     Multiplexable, locus-specific targeting of long RNAs with 
            CRISPR-Display.
  JOURNAL   Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 
            2015 Jun 1.
  PUBMED    26030444
REFERENCE   2  (bases 1 to 1050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.3433"; journalName: "Nat Methods"; date: "2015-07"; 
            volume: "12"; issue: "7"; pages: "664-70"
COMMENT     Sequence Label: pU6_(GLuc)_sgRNA
FEATURES             Location/Qualifiers
     source          1..1050
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        45..120
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(189..205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(189..205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(202..224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    213..229
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(237..243,244..261,262..267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    282..303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(420..437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(671..690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcttgtggaa aggacgaaac accgatctag atacgactca ctatgtttta gagctagaaa
       61 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      121 tttttttctt aattaagtct agagtcgact gtttaaacct gcaggcatgc aagcttggcg
      181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1021 tacactagaa gaacagtatt tggtatctgc
//
